LOCUS       40924_44978        6078 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18-mini-Tn7T-Gm-Gateway, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6078)
  AUTHORS   Choi K-H., Gaynor JB, White KG, Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     A Tn7-based universal bacterial cloning and expression system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6078)
  AUTHORS   Choi K-H., Gaynor JB, White KG, Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-AUG-2004) Microbiology, Immunology and Pathology, 
            Colorado State University, Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 6078)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6078)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-AUG-2004) Microbiology, Immunology and Pathology, Colorado State
            University, Fort Collins, CO 80523, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   382..580
                     /label=Tn7R, right end of Tn7
                     /note="Tn7R, right end of Tn7"
     primer_bind     585..601
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      629..723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      826..912
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    958..1005
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1138..1668)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(1857..1885)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    1926..1973
                     /label=FRT, Flp recombinase target site
                     /note="FRT, Flp recombinase target site"
     protein_bind    1926..1973
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    2089..2213
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2257..2559)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(2904..3560)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3614..3644)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(3669..3793)
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     mobile_element  complement(3824..3989)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      complement(4259..4847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5021..5878)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5879..5983)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct agaggaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
      361 tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga
      421 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
      481 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
      541 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ggttcgaggt cgacggtatc
      601 gataagctag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
      661 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
      721 aatccaagct agactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
      781 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
      841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      901 agtaggacaa atccgccgct aggagcttgc ggcccggacg atcgaattgg ggatcttgaa
      961 gtacctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag
     1021 ctgatgtgct taaaaactta ctcaatggaa ttctagatcg aattggccgc ggcgttgtga
     1081 caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt
     1141 ggcggtactt gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata
     1201 gagagccact gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag
     1261 cgcgttggcc tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg
     1321 ctcgccggag actgcgagat catagatata gatctcacta cgcggctgct caaacttggg
     1381 cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat
     1441 gaatgtctta ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta
     1501 ggtggctacg tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac
     1561 ttggtcaggg ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt
     1621 tttagggcga ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc
     1681 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     1741 actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc
     1801 tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt
     1861 tacgaaccga acaggcttat gtcaattcga tccattgctg ttgacaaagg gaatcagggg
     1921 atcttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttcagagcgc
     1981 ttttgaagct aattcgatca tgcatgagct cactagtgga tcccccgggc tgcaggaatt
     2041 cctcgagaag cttgcatgcc tgcaggtcga ctctagagga tccccatcac cactttgtac
     2101 aagaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg
     2161 cataaaaaac agactacata atactgtaaa acacaacata tccagtcact atggtcgacc
     2221 tgcagactgg ctgtgtataa cggagcctga catttatatt ccccagaaca tcaggttaat
     2281 ggcgtttttg atgtcatttt cgcggtggct gagatcagcc acttcttccc cgataacgga
     2341 gaccggcaca ctggccatat cggtggtcat catgcgccag ctttcatccc cgatatgcac
     2401 caccgggtaa agttcacggg agactttatc tgacagcaga cgtgcactgg ccagggggat
     2461 caccatccgt cgcccgggcg tgtcaataat atcactctgt acatccacaa acagacgata
     2521 acggctctct cttttatagg tgtaaacctt aaactgcatt tcaccagtcc ctgttctcgt
     2581 cagcaaaaga gccgttcatt tcaataaacc gggcgacctc agccatccct tcctgatttt
     2641 ccgctttcca gcgttcggca cgcagacgac gggcttcatt ctgcatggtt gtgcttacca
     2701 gaccggagat attgacatca tatatgcctt gagcaactga tagctgtcgc tgtcaactgt
     2761 cactgtaata cgctgcttca tagcacacct ctttttgaca tacttcgggt atacatatca
     2821 gtatatattc ttataccgca aaaatcagcg cgcaaatacg catactgtta tctggctttt
     2881 agtaagccgg atccacgcgt ttacgccccg ccctgccact catcgcagta ctgttgtaat
     2941 tcattaagca ttctgccgac atggaagcca tcacagacgg catgatgaac ctgaatcgcc
     3001 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     3061 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     3121 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     3181 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     3241 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     3301 ctatcccata tcaccagctc accgtctttc attgccatac ggaattccgg atgagcattc
     3361 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     3421 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     3481 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     3541 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcgc cggatcctaa
     3601 ctcaaaatcc acacattata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3661 gcggccgcca tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat
     3721 gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt tcagcttttt
     3781 tgtacaaact tgtgatgggt acctcgcgaa ggccttgcag gccaaccaga taagtgaaat
     3841 ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg ctacacccag
     3901 ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt tccacccctc
     3961 ccagttccca actattttgt ccgcccacag cggggcattt ttcttcctgt tatgtttggg
     4021 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4081 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4141 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4201 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4261 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4321 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4381 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4441 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4501 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4561 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4621 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4681 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4741 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4801 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4861 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4921 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4981 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5041 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5101 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5161 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5221 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5281 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5341 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5401 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5461 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5521 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5581 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5641 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5701 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5761 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5821 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5881 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5941 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6001 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6061 tcacgaggcc ctttcgtc
//
